宫仁梅, 万超群, 任红宇, 卢锡华. 军团菌16SrRNA聚合酶链反应方法的建立[J]. 疾病监测, 1996, 11(2): 68-71.
引用本文: 宫仁梅, 万超群, 任红宇, 卢锡华. 军团菌16SrRNA聚合酶链反应方法的建立[J]. 疾病监测, 1996, 11(2): 68-71.
Gong Shimei et al, . Establishment and Preliminary Application of a 16SrRNA Polymerase Chain Reaction Assay for Detection of Legionella Species[J]. Disease Surveillance, 1996, 11(2): 68-71.
Citation: Gong Shimei et al, . Establishment and Preliminary Application of a 16SrRNA Polymerase Chain Reaction Assay for Detection of Legionella Species[J]. Disease Surveillance, 1996, 11(2): 68-71.

军团菌16SrRNA聚合酶链反应方法的建立

Establishment and Preliminary Application of a 16SrRNA Polymerase Chain Reaction Assay for Detection of Legionella Species

  • 摘要: 本文建立了一个用于检测军团菌的多聚酶链反应方法。扩增参考菌株的染色体DNA(L.pneumophital-14、L.micdadei、L.dumoffii Llongbeachae、L.Jordanis、L.bozemanii),均可检出375bp的16SrRNA基因片段。对于已经监定的5株国内分离菌株染色体DNA进行扩增,获得了相同的结果,地高辛标记16SrRNA基因探针与扩增产物杂交,结果为阳性。而扩增14株非军团菌均为阴性。采用煮沸法制备细菌染色体DNA,PCR法检测环境水军团菌敏感性为280cfu/ml水,检查临床标本军团菌为560cfu/ml支气管灌洗液。上述结果表明该法敏感、快速、简便,具有较好的特异性。

     

    Abstract: A 165rRNA polymerasse chain reaction CPCR) assay for the detection of Le- gionella Species was established. A 375 bp fragment of the 16SrRNA gene could be detected by amplifing genomic DNA of reference strains (Lp1-14, Lm, L1, Ld, Lb, Lg). The same result was also obtained by amplifing 5 identified strains isolated from China. The amplified products were verified by hybridization with 16SrRNA gene probes. However, 14 non-legionella strains could't produced a positive amplified fragment. Preparing bacterial genomic DNA by boiling method,the assay could detect 280cfu/ml water and 560 cfu/ml clinical bronchial fluid respectively. Above- mentioned result showed that this assay was sensitive、rapid、specific and easy to perform.

     

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